18 resultados para label-free quantification
em Universidade do Minho
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Supplementary data associated with this article can be found, in the online version, at: http://dx.doi.org/10.1016/j.electacta.2015.09.169.
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Injectable biomaterials with in situ cross-linking reactions have been suggested to minimize the invasiveness associated with most implantation procedures. However, problems related with the rapid liquid-to-gel transition reaction can arise because it is difficult to predict the reliability of the reaction and its end products, as well as to mitigate cytotoxicity to the surrounding tissues. An alternative minimally invasive approach to deliver solid implants in vivo is based on injectable microparticles, which can be processed in vitro with high fidelity and reliability, while showing low cytotoxicity. Their delivery to the defect can be performed by injection through a small diameter syringe needle. We present a new methodology for the continuous, solvent- and oil-free production of photopolymerizable microparticles containing encapsulated human dermal fibroblasts. A precursor solution of cells in photo-reactive PEG-fibrinogen (PF) polymer was transported through a transparent injector exposed to light-irradiation before being atomized in a jet-in-air nozzle. Shear rheometry data provided the cross-linking kinetics of each PF/cell solution, which was then used to determine the amount of irradiation required to partially polymerize the mixture prior to atomization. The partially polymerized drops fell into a gelation bath for further polymerization. The system was capable of producing cell-laden microparticles with high cellular viability, with an average diameter of between 88.1 µm to 347.1 µm and a dispersity of between 1.1 and 2.4, depending on the parameters chosen.
Tendon regeneration through a scaffold-free approach: development of tenogenic magnetic hASCs sheets
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Tendon's regeneration is limited, demanding for cell-based strategies to fully restore their functionality upon injury. The concept of magnetic force-based TE(1), generally using magnetic nanoparticles may enable, for example, stem cell stimulation and/or remote control over TE constructs. Thus, we originally propose the development of magnetic cell sheets (magCSs) with tenogenic capability, aimed at promoting tendon's regeneration. A Tenomodulin (TNMD+) subpopulation was sorted from human adipose stem cells (hASCs), using TNMD-coated immunomagnetic beads(2) and used as cell source for the development of magCSs. Briefly, cells were labeled with iron oxide composite particles (Micromod) and cultured for 7 days in α-MEM medium with or without magnetic stimulation provided by a magnetic device (nanoTherics). CSs were retrieved from the plates using magnet attraction as contiguous sheets of cells within its own deposited ECM.
Connecting free volume with shape memory properties in noncytotoxic gamma-irradiated polycyclooctene
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The free volume holes of a shape memory polymer have been analysed considering that the empty space between molecules is necessary for the molecular motion, and the shape memory response is based on polymer segments acting as molecular switches through variable flexibility with temperature or other stimuli. Therefore, thermomechanical analysis (TMA) and positron annihilation lifetime spectroscopy (PALS) have been applied to analyse shape recovery and free volume hole sizes in gamma irradiated polycyclooctene (PCO) samples, as a non-cytotoxic alternative to more conventional PCO crosslinked via peroxide for future applications in medicine. Thus, a first approach relating structure, free volume holes and shape memory properties in gamma irradiated PCO is presented. The results suggest that free volume holes caused by gamma irradiation in PCO samples facilitate the recovery process by improving movement of polymer chains and open t possibilities for the design and control of the macroscopic response.
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In some regions of Brazil, especially where the water is scarce, drinking water is stored in water storage tanks. This practice gives the consumer the guarantee of available water. The water storage conditions such as the exposure to hot weather when the tanks are on rooftops allow the development of microorganisms and microbial biofilms which can deteriorate the water quality and increase the risk to human health [1,2]. This study describes the filamentous fungi (FF) detected in free water and biofilms in drinking water storage tanks in Recife - Pernambuco, Brazil. Five sampling times in triplicate were performed at two distinct points. Colony-forming units (CFU) of FF fungi were determined with 0.45 µm filtration membranes using peptone glucose rose Bengal agar (PGRBA). From the 30 samples analysed a total of 1136 CFU were obtained. The water biofilms were collected from samplers consisting of polyethylene coupons, previously installed in the reservoirs. These coupons were transferred to PGRBA plates and incubated using with the same conditions described for free FF. For the in situ detection of FF in biofilms the Calcofluor White staining technique was used. This procedure demonstrated FF forming biofilms on the surfaces of the coupons. Brazilian legislation does not define limits for FF in drinking water. However considering the potential risk of fungal contamination, the data obtained in this study will contribute to developing future quantitative and qualitative parameters for the presence of fungi in drinking water distribution systems in Brazil. [1] HageskaL, G, Lima, N, Skaar, I. The study of fungi in drinking water. Mycological Research, 113, 2009, 165-172. [2] Skaar I, Hageskal G. Fungi in Drinking Water. In.: Paterson RRM, Lima N. (Eds.) Molecular Biology of Food and Water Borne Mycotoxigenic and Mycotic Fungi. CRC Press, Taylor & Francis Group, Boca Raton, 2015, 597-606.
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Rational manipulation of mRNA folding free energy allows rheostat control of pneumolysin production by Streptococcus pneumoniae
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Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Biomateriais, Reabilitação e Biomecânica)
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O sector do calçado português apresenta excelentes indicadores económicos e um desempenho competitivo assinalável. Os últimos quatro anos foram particularmente fortes, tendo Portugal atingido o 11º lugar entre os 15 maiores exportadores mundiais, com o segundo preço mais elevado por par de sapato vendido, superado apenas pela Itália. O artigo analisa o tipo de inovação realizada pelas empresas consideradas inovadoras dentro do sector do calçado em Portugal, de acordo com o manual de Oslo, e analisa a evolução que as empresas fizeram ao passarem de um regime dominado pelo private label para o desenvolvimento de produtos, de coleções e de marcas próprias. A pesquisa foi de natureza qualitativa, envolvendo sete empresas com inovação, sendo realizado estudos de caso através de entrevistas semi-estruturadas, análise de documentos e visitas às empresas e aos seus processos produtivos e de criação. Os resultados obtidos na investigação evidenciam uma relação entre os indicadores económicos e de desempenho e a existência de determinadas categorias de inovação. Evidencia ainda a relação existente entre o regime de produção e os indicadores económicos das empresas.
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Tese de Doutoramento em Biologia de Plantas
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Architectural (bad) smells are design decisions found in software architectures that degrade the ability of systems to evolve. This paper presents an approach to verify that a software architecture is smellfree using the Archery architectural description language. The language provides a core for modelling software architectures and an extension for specifying constraints. The approach consists in precisely specifying architectural smells as constraints, and then verifying that software architectures do not satisfy any of them. The constraint language is based on a propositional modal logic with recursion that includes: a converse operator for relations among architectural concepts, graded modalities for describing the cardinality in such relations, and nominals referencing architectural elements. Four architectural smells illustrate the approach.
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In some regions of Brazil, especially where the water is scarce, drinking water is stored in water storage tanks. This practice gives the consumer the guarantee of available water. The water storage conditions such as the exposure to hot weather when the tanks are on rooftops allow the development of microorganisms and microbial biofilms which can deteriorate the water quality and increase the risk to human health [1,2]. This study describes the filamentous fungi (FF) detected in free water and biofilms in drinking water storage tanks in Recife - Pernambuco, Brazil. Five sampling times in triplicate were performed at two distinct points. Colony-forming units (CFU) of FF fungi were determined with 0.45 μm filtration membranes using peptone glucose rose Bengal agar (PGRBA). From the 30 samples analysed a total of 1136 CFU were obtained. The water biofilms were collected from samplers consisting of polyethylene coupons, previously installed in the reservoirs. These coupons were transferred to PGRBA plates and incubated using with the same conditions described for free FF. For the in situ detection of FF in biofilms the Calcofluor White staining technique was used. This procedure demonstrated FF forming biofilms on the surfaces of the coupons. Brazilian legislation does not define limits for FF in drinking water. However considering the potential risk of fungal contamination, the data obtained in this study will contribute to developing future quantitative and qualitative parameters for the presence of fungi in drinking water distribution systems in Brazil.
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Dissertação de mestrado em Ciências da Comunicação (área de especialização em Publicidade e Relações Públicas)
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Dissertação de mestrado em Biofísica e Bionanossistemas
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Fluorescence in situ hybridization (FISH) is based on the use of fluorescent staining dyes, however, the signal intensity of the images obtained by microscopy is seldom quantified with accuracy by the researcher. The development of innovative digital image processing programs and tools has been trying to overcome this problem, however, the determination of fluorescent intensity in microscopy images still has issues due to the lack of precision in the results and the complexity of existing software. This work presents FISHji, a set of new ImageJ methods for automated quantification of fluorescence in images obtained by epifluorescence microscopy. To validate the methods, results obtained by FISHji were compared with results obtained by flow cytometry. The mean correlation between FISHji and flow cytometry was high and significant, showing that the imaging methods are able to accurately assess the signal intensity of fluorescence images. FISHji are available for non-commercial use at http://paginas.fe.up.pt/nazevedo/.